Biotinyl-l-3-(2-naphthyl)-alanine hydrazide derivatives of N-glycans: versatile solid-phase probes for carbohydrate-recognition studies.
Biotinyl-oligosaccharides are a relatively new generation of saccharide probes that enable immobilization of desired oligosaccharides on streptavidin matrices for studies of carbohydrate-protein interactions. Here we describe the facile preparation of biotinyl-l-3-(2-naphthyl)-alanine hydrazide (BNAH) derivatives of oligosaccharides, containing a strong UV absorbing and fluorescent group, in which the ring of the reducing-end monosaccharide is nonreduced. We evaluate reactivities of immobilized BNAH- N -glycans with plant lectins that recognize aspects of the oligosaccharide core or outer-arms. We make some comparisons with 2-amino-6-amidobiotinyl-pyridine (BAP) derivatives obtained by reductive amination, and 6-(biotinyl)-aminocaproyl-hydrazide (BACH) derivatives which have a longer spacer-arm. N -Glycan-BNAH and-BAP derivatives have, overall, comparable reactivities with lectins which recognize N -glycan outer-arms or the trimannosyl core, but only BNAH and BACH derivatives are bound by lectins which recognize the non-reduced core. Moreover, with Pisum sativum agglutinin (PSA) which additionally requires the fucosyl- N- glycan-asparaginyl core for high affinity binding, the immobilized BNAH derivative (which is an alanine hydrazide beta-glycoside) can substitute for the natural beta-glycosylasparaginyl core, whereas the BACH derivative (aminocaproyl-hydrazide-beta-glycoside) is less effective. BNAH is a derivatization reagent of choice, therefore, for solid phase carbohydrate-binding experiments with immobilized N -glycans.
Leteux
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The Glycosciences Laboratory, Imperial College School of Medicine, Harrow, Middlesex, United Kingdom.
Childs
R A
RA
Chai
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Stoll
M S
MS
Kogelberg
H
H
Feizi
T
T
D000371:Agglutination / Q000187:drug effects; D000409:Alanine / Q000031:analogs & derivatives; D001710:Biotin / Q000031:analogs & derivatives; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D004912:Erythrocytes / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D006834:Hydrazines / Q000737:chemistry; D037102:Lectins / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009281:Naphthalenes / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*; D018532:Pisum sativum / Q000737:chemistry; D037121:Plant Lectins; D010940:Plant Proteins / Q000378:metabolism; D011485:Protein Binding; D019809:Streptavidin / Q000378:metabolism
D005456:Fluorescent Dyes; D006834:Hydrazines; D037102:Lectins; D009281:Naphthalenes; D009844:Oligosaccharides; D037121:Plant Lectins; D010940:Plant Proteins; D001710:Biotin; D019809:Streptavidin; D000409:Alanine